Inhibition Of Fast Axonal Transport Research Articles

Molecular Determinants of the Neurotoxicity of Acrylamide Through Covalent Docking

Acrylamide is formed during food processing by Maillard reaction between sugars and proteins at high temperatures. Besides, acrylamide is used in many industries, from water waste treatment to manufacture of paper, fabrics, dyes or cosmetics. Cumulative exposure to acrylamide, either from diet or in the workplace, may result in neurotoxicity, involving three possible mechanisms: inhibition of fast axonal transport, alteration of neurotransmitter levels, and direct inhibition of neurotransmission. From the molecular point of view, acrylamide may affect protein function by forming a covalent adduct with cysteine residues via a Michael addition reaction. However, the molecular determinants of acrylamide reactivity are not completely understood and further investigation is needed. As a follow-up of previous computational studies by Carloni and coworkers [1,2], here we have performed a systematic study of the acrylamide protein targets reported so far in the literature in connection with neurotoxicity using covalent docking. Comparison with drug protein targets of acrylamide warhead-containing inhibitors was also performed. Analysis of the acrylamide-protein complexes provides insights into the determinants of cysteine reactivity, as well as the residue composition of acrylamide binding sites. This information could be used to predict other potential protein targets mediating acrylamide neurotoxicity. This project was inspired by late Dr. Ernesto Illy and funded by the Ernesto Illy Foundation. [1] de Lima, E. F., & Carloni, P. (2011). Acrylamide Binding to Its Cellular Targets: Insights from Computational Studies. Computational Biology and Applied Bioinformatics, 431. [2] Papamokos, G., Dreyer, J., Navarini, L., & Carloni, P. (2014). Trapping acrylamide by a Michael addition: a computational study of the reaction between acrylamide and niacin. Int. J. Quantum Chem., 114(9), 553-559.

A MultiTEP platform-based epitope vaccine targeting the phosphatase activating domain (PAD) of tau: therapeutic efficacy in PS19 mice

Pathological tau correlates well with cognitive impairments in Alzheimer’s disease (AD) patients and therefore represents a promising target for immunotherapy. Targeting an appropriate B cell epitope in pathological tau could in theory produce an effective reduction of pathology without disrupting the function of normal native tau. Recent data demonstrate that the N-terminal region of tau (aa 2-18), termed the “phosphatase activation domain (PAD)”, is hidden within native Tau in a ‘paperclip’-like conformation. Conversely, PAD is exposed in pathological tau and plays an essential role in the inhibition of fast axonal transport and tau polymerization. Thus, we hypothesized that anti-tau2-18 antibodies may safely and specifically reduce pathological tau and prevent further aggregation, which in turn would neutralize tau toxicity. Therefore, we evaluated the immunogenicity and therapeutic efficacy of our MultiTEP platform-based vaccine targeting tau2-18 formulated with AdvaxCpG adjuvant (AV-1980R/A) in PS19 tau transgenic mice. The AV-1980R/A induced extremely high antibody responses and the resulting sera recognized neurofibrillary tangles and plaque-associated dystrophic neurites in AD brain sections. In addition, under non-denaturing conditions AV-1980R/A sera preferentially recognized AD-associated tau. Importantly, vaccination also prevented age-related motor and cognitive deficits in PS19 mice and significantly reduced insoluble total and phosphorylated tau species. Taken together, these findings suggest that predominantly targeting misfolded tau with AV-1980R/A could represent an effective strategy for AD immunotherapy.

Protein Misfolding, Signaling Abnormalities and Altered Fast Axonal Transport: Implications for Alzheimer and Prion Diseases.

Histopathological studies revealed that progressive neuropathies including Alzheimer, and Prion diseases among others, include accumulations of misfolded proteins intracellularly, extracellularly, or both. Experimental evidence suggests that among the accumulated misfolded proteins, small soluble oligomeric conformers represent the most neurotoxic species. Concomitant phenomena shared by different protein misfolding diseases includes alterations in phosphorylation-based signaling pathways synaptic dysfunction, and axonal pathology, but mechanisms linking these pathogenic features to aggregated neuropathogenic proteins remain unknown. Relevant to this issue, results from recent work revealed inhibition of fast axonal transport (AT) as a novel toxic effect elicited by oligomeric forms of amyloid beta and cellular prion protein PrPC, signature pathological proteins associated with Alzheimer and Prion diseases, respectively. Interestingly, the toxic effect of these oligomers was fully prevented by pharmacological inhibitors of casein kinase 2 (CK2), a remarkable discovery with major implications for the development of pharmacological target-driven therapeutic intervention for Alzheimer and Prion diseases.

Prion protein inhibits fast axonal transport through a mechanism involving casein kinase 2.

Prion diseases include a number of progressive neuropathies involving conformational changes in cellular prion protein (PrPc) that may be fatal sporadic, familial or infectious. Pathological evidence indicated that neurons affected in prion diseases follow a dying-back pattern of degeneration. However, specific cellular processes affected by PrPc that explain such a pattern have not yet been identified. Results from cell biological and pharmacological experiments in isolated squid axoplasm and primary cultured neurons reveal inhibition of fast axonal transport (FAT) as a novel toxic effect elicited by PrPc. Pharmacological, biochemical and cell biological experiments further indicate this toxic effect involves casein kinase 2 (CK2) activation, providing a molecular basis for the toxic effect of PrPc on FAT. CK2 was found to phosphorylate and inhibit light chain subunits of the major motor protein conventional kinesin. Collectively, these findings suggest CK2 as a novel therapeutic target to prevent the gradual loss of neuronal connectivity that characterizes prion diseases.

Humanized monoclonal antibody armanezumab specific to N-terminus of pathological tau: characterization and therapeutic potency

BackgroundThe experience from clinical trials indicates that anti-Aβ immunotherapy could be effective in early/pre-clinical stages of AD, whereas at the late stages promoting the clearing of Aβ alone may be insufficient to halt the disease progression. At the same time, pathological tau correlates much better with the degree of dementia than Aβ deposition. Therefore, targeting pathological tau may provide a more promising approach for the treatment of advanced stages of AD. Recent data demonstrates that the N-terminal region of tau spanning aa 2–18 termed “phosphatase activation domain” that is normally hidden in the native protein in ‘paperclip’-like conformation, becomes exposed in pathological tau and plays an essential role in the inhibition of fast axonal transport and in aggregation of tau. Hence, we hypothesized that anti-Tau2–18 monoclonal antibodies (mAb) may recognize pathological, but not normal tau at very early stages of tauopathy and prevent or decrease the aggregation of this molecule.MethodsMouse mAbs were generated using standard hybridoma methodology. CDR grafting was used for humanization of mouse mAb. Humanized mAb (Armanezumab) was characterized and tested in vitro/ex vivo/in vivo using biochemical and immunological methods (HPLC, Biacore, ELISA, IHC, FRET, etc.). Stable DG44 cell line expressing Armanezumab was generated by clone selection with increased concentrations of methotrexate (MTX).ResultsA panel of mouse mAbs was generated, clone 1C9 was selected based on binding to pathological human tau with high affinity and humanized. Fine epitope mapping revealed conservation of the epitope of human tau recognized by the parent murine mAb and Armanezumab. Importantly, Armanezumab (i) bound to tau with high affinity as determined by Biacore; (ii) bound pathological tau in brains from AD, FTD and Pick’s disease cases; (iii) inhibited seeding effect of aggregated tau from brain lysate of P301S Tg mice; (iv) inhibited cytotoxic effect of tau oligomers; (v) reduced total tau (HT7) and AT100, PHF1, AT8, AT180, p212, p214-positive tau species in brains of tau transgenic mice after intracranial injection. A stable CHO cell line producing >1.5 g/l humanized mAb, Armanezumab was generated.ConclusionThese findings suggest that Armanezumab could be therapeutic in clinical studies for treatment of AD.

Inhibition of fast axonal transport by pathogenic SOD1 involves activation of p38 MAP kinase.

Dying-back degeneration of motor neuron axons represents an established feature of familial amyotrophic lateral sclerosis (FALS) associated with superoxide dismutase 1 (SOD1) mutations, but axon-autonomous effects of pathogenic SOD1 remained undefined. Characteristics of motor neurons affected in FALS include abnormal kinase activation, aberrant neurofilament phosphorylation, and fast axonal transport (FAT) deficits, but functional relationships among these pathogenic events were unclear. Experiments in isolated squid axoplasm reveal that FALS-related SOD1 mutant polypeptides inhibit FAT through a mechanism involving a p38 mitogen activated protein kinase pathway. Mutant SOD1 activated neuronal p38 in mouse spinal cord, neuroblastoma cells and squid axoplasm. Active p38 MAP kinase phosphorylated kinesin-1, and this phosphorylation event inhibited kinesin-1. Finally, vesicle motility assays revealed previously unrecognized, isoform-specific effects of p38 on FAT. Axon-autonomous activation of the p38 pathway represents a novel gain of toxic function for FALS-linked SOD1 proteins consistent with the dying-back pattern of neurodegeneration characteristic of ALS.

Huntington-Associated Phosphorylation of Kinesin-1 Enhances Autoinhibition in a Phosphomimic

One of the consequences of the triplet expansion in Huntington's disease is inhibition of fast axonal transport (FAT). Phosphorylation of Ser176 in human kinesin-1 by JNK3 has been implicated in this inhibition (Morfini, et al., Nature Neruo. 12, 866 (2009)). To investigate the molecular basis for the inhibition of FAT, we have generated the S182E phosphomimic of the homologous residue in Drosophila kinesin-1. When introduced into short dimer of motor domains, the S182E mutation produces a 30% decrease in the maximum rate of microtubule-stimulated ATPase rate in solution and a similar reduction in the sliding rate of axonemes in a multimotor sliding assay. This only modest decrease suggests that direct inhibition of motility is not likely to be the principal cause of the pronounced inhibition of FAT. However, free kinesin is known to be autoinhibited through the binding of a tail domain to a dimer of motor domains (heads) and the Ser182 phosphorylation site is near the tail binding site on the heads (Kaan, et al., Science 333, 883 (2011)) where it could influence autoinhibition. One possibility is that the increased negative charge on the heads due to phosphorylation of Ser182 could produce a stronger interaction with the positively charge tail domain that would strengthen autoinhibition and inhibit FAT. To test the effect of the phosphomimic on autoinhibition, the binding of a monomeric tail domain to a dimer of motor domains was determined using a FRET assay. In 100 mM KCl, monomeric tail domains bind to mutant S182E heads three-fold more tightly than to wild type heads. This suggest that inhibition of FAT may principally be due to enhanced tail binding and accompanying autoinhibition of kinesin-1 following phosphorylation.Supported by NIH Grant NS058848.

Chronic, Intermittent Exposure to Chlorpyrifos in Rats: Protracted Effects on Axonal Transport, Neurotrophin Receptors, Cholinergic Markers, and Information Processing

Persistent behavioral abnormalities have been commonly associated with acute organophosphate (OP) pesticide poisoning; however, relatively little is known about the consequences of chronic OP exposures that are not associated with acute cholinergic symptoms. In this study, the behavioral and neurochemical effects of chronic, intermittent, and subthreshold exposures to the OP pesticide, chlorpyrifos (CPF), were investigated. Rats were injected with CPF s.c. (dose range, 2.5-18.0 mg/kg) every other day over the course of 30 days and then were given a 2-week CPF-free washout period. In behavioral experiments conducted during the washout period, dose-dependent decrements in a water-maze hidden platform task and a prepulse inhibition procedure were observed, without significant effects on open-field activity, Rotorod performance, grip strength, or a spontaneous novel object recognition task. After washout, levels of CPF and its metabolite 3,5,6-trichloro-2-pyridinol were minimal in plasma and brain; however, cholinesterase inhibition was still detectable. Furthermore, the 18.0 mg/kg dose of CPF was associated with (brain region-dependent) decreases in nerve growth factor receptors and cholinergic proteins including the vesicular acetylcholine transporter, the high-affinity choline transporter, and the alpha(7)-nicotinic acetylcholine receptor. These deficits were accompanied by decreases in anterograde and retrograde axonal transport measured in sciatic nerves ex vivo. Thus, low-level (intermittent) exposure to CPF has persistent effects on neurotrophin receptors and cholinergic proteins, possibly through inhibition of fast axonal transport. Such neurochemical changes may lead to deficits in information processing and cognitive function.

JNK mediates pathogenic effects of polyglutamine-expanded androgen receptor on fast axonal transport

Expansion of the polyglutamine (polyQ) stretch in the androgen receptor (AR) protein leads to spinal and bulbar muscular atrophy (SBMA), a neurodegenerative disease characterized by lower motor neuron degeneration. The pathogenic mechanisms underlying SBMA remain unknown, but recent experiments show that inhibition of fast axonal transport (FAT) by polyQ-expanded proteins, including polyQ-AR, represents a new cytoplasmic pathogenic lesion. Using pharmacological, biochemical and cell biological experiments, we found a new pathogenic pathway that is affected in SBMA and results in compromised FAT. PolyQ-AR inhibits FAT in a human cell line and in squid axoplasm through a pathway that involves activation of cJun N-terminal kinase (JNK) activity. Active JNK phosphorylated kinesin-1 heavy chains and inhibited kinesin-1 microtubule-binding activity. JNK inhibitors prevented polyQ-AR-mediated inhibition of FAT and reversed suppression of neurite formation by polyQ-AR. We propose that JNK represents a promising target for therapeutic interventions in SBMA.

Coexpression of GSK-3β Corrects Phenotypic Aberrations of Dorsal Root Ganglion Cells, Cultured from Adult Transgenic Mice Overexpressing Human Protein tau

Coexpression of constitutively active GSK-3β[S9A] rescued the axonal pathology induced by overexpression of human tau in transgenic mice (Spittaels et al., (2000) J. Biol. Chem. 275, 41340–41349). We isolated dorsal root ganglion (DRG) neuronal cultures from adult tau4R- and tau4R × GSK-3β-transgenic mice to define the mechanisms at the cellular and subcellular level. DRG from tau4R-transgenics showed a reduced sprouting capacity while density and stability of microtubules in the axonal processes were significantly increased. Video-enhanced contrast microscopy demonstrated a dramatic inhibition of fast axonal transport. Coexpression of GSK-3β increased tau phosphorylation and reversed the effects on microtubule stability and saltatory motion. In DRG from GSK-3β single transgenics, increased tau phosphorylation was evident without any major effects on microtubule stability or axonal transport. These observations support the hypothesis that excess tau competed with motor-proteins for binding to microtubules and/or that a rigid microtubular system inhibits axonal transport.

Fast axonal transport is required for growth cone advance.

Growth cones are capable of advancing despite linkage to a stationary axonal cytoskeleton in chick and murine dorsal root ganglion neurites. Several lines of evidence point to the growth cone as the site of cytoskeletal elongation. Fast axonal transport is probably the means by which cytoskeletal elements or cofactors are rapidly moved through the axon. We report that direct, but reversible, inhibition of fast axonal transport with laser optical tweezers inhibits growth cone motility if cytoskeletal attachment to the cell body is maintained. Advancement ceases after a distance-dependent lag period which correlates with the rate of fast axonal transport. But severing the axonal cytoskeleton with the laser tweezers allows growth cones to advance considerably further. We suggest that axon elongation requires fast axonal transport but growth cone motility does not.

A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm.

One of our monoclonal antibodies against the heavy chain of bovine kinesin (H2) also recognized the heavy chain of squid kinesin. The immunofluorescence pattern of H2 in axoplasm was similar to that seen in mammalian cells with antibodies specific for kinesin light and heavy chains, indicating that squid kinesin is also concentrated on membrane-bounded organelles. Although kinesin is assumed to be a motor for translocation of membrane-bounded organelles in fast axonal transport, direct evidence has been lacking. Perfusion of axoplasm with purified H2 at 0.1-0.4 mg/ml resulted in a profound inhibition of both the rates and number of organelles moving in anterograde and retrograde directions in the interior of the axoplasm, and comparable inhibition was noted in bidirectional movement along individual microtubules at the periphery. Maximal inhibition developed over 30-60 min. Perfusion with higher concentrations of H2 (greater than 1 mg of IgG per ml) were less effective, whereas perfusion with 0.04 mg of H2 per ml resulted in minimal inhibition. Movement of membrane-bounded organelles after perfusion with comparable levels of irrelevant mouse IgG (0.04 to greater than 1 mg/ml) were not distinguishable from perfusion with buffer controls. Inhibition of fast axonal transport by an antibody specific for kinesin provides direct evidence that kinesin is involved in the translocation of membrane-bounded organelles in axons. Moreover, the inhibition of bidirectional axonal transport by H2 raises the possibility that kinesin may play some role in both anterograde and retrograde axonal transport.

Mechanisms of the inhibition of fast axonal transport by local anesthetics

The present study attempted to clarify the mechanism(s) by which local anesthetics inhibit fast axonal transport. Spinal nerves of the bullfrog were incubated with local anesthetics under conditions known to inhibit transport and the effects of these exposures to local anesthetics on the content of adenosine triphosphate and creatine phosphate in nerves and on the density of microtubules in unmyelinated axons were examined. Lidocaine, at concentrations of 14 or 20 mM, did not reduce significantly the content of adenosine triphosphate (although significant reductions in creatine phosphate were observed); the density of mierotubules was also not affected by 14 mM lidocaine. Some mechanism other than inhibition of oxidative metabolism or disruption of microtubules must therefore be responsible for the inhibition of fast axonal transport by 14 mM lidocaine. Significant reductions in the content of adenosine triphosphate were observed with 1 or 2 mM tetracaine and with 0.5 or 1 mM dibucaine (this latter concentration of dibucaine also reduced the content of creatine phosphate); however, comparison with the effects of 2,4-dinitrophenol indicated that these inhibitions of oxidative metabolism were insufficient to inhibit transport in the case of 0.5mM dibucaine or could at best only partly explain the inhibition of transport in the other cases. Since the density of microtubules was not affected by 1 mM tetracaine and was not sufficiently reduced by 0.5 mM dibucaine to inhibit transport, some other effect must again largely contribute to or be solely responsible for the inhibition of fast axonal transport by these concentrations of dibucaine and tetracaine.

Dissociation of the inhibition of fast axonal transport by chlorimipramine from an effect on axonal microtubules

The effect of in vitro exposure of bullfrog spinal nerves to 0.2 mM chlorimipramine on the density of axonal microtubules was studied in an attempt to clarify the mechanism by which chlorimipramine inhibits fast axonal transport. A 17-h exposure to chlorimipramine reduced the density of microtubules in unmyelinated axons by only 18%; this microtubular loss does not reach the upper limit of the range of microtubule reduction associated with inhibition of fast axonal transport. A 23-h exposure to chlorimipramine, which had decreased microtubular density in unmyelinated axons by 40% in a previous study, did not decrease microtubular density in myelinated axons in the present study. These results rule out microtubular destruction as the mechanism responsible for inhibition of fast orthograde axonal transport by chlorimipramine, and greatly reduce the likelihood that microtubular destruction plays a significant role in the inhibition of fast retrograde transport by chlorimipramine.

Pressure-induced inhibition of fast axonal transport of proteins in the rabbit vagus nerve in galactose neuropathy: prevention by an aldose reductase inhibitor

Fast and slow anterograde axonal transport and retrograde axonal transport of proteins were studied in the mainly non-myelinated sensory fibres of the vagus nerve of rabbits fed a diet of 50% galactose over a period of 29 days. Galactose feeding had no effect on the rate or protein composition of slow transport nor on the amount of retrogradely transported proteins. There was a slight retardation of fast transported proteins although their composition was unchanged. The galactose feeding led to a significant increase (p less than 0.005) in nerve water content and nerve galactitol but no significant change in myo-inositol. When 20 mm Hg pressure was applied locally to the cervical vagus nerve, fast transported proteins accumulated proximal to the compression zone in the galactose-fed but not in control rabbits. Administration of the aldose reductase inhibitor Statil (ICI 128436) throughout the experiment prevented the increased susceptibility to pressure and the increase in nerve galactitol and water content. The effects of pressure are similar to those found in the streptozotocin-diabetic rat although the underlying mechanisms may differ.

Treatment with an aldose reductase inhibitor can reduce the susceptibility of fast axonal transport following nerve compression in the streptozotocin-diabetic rat

The effect of treatment with an aldose reductase inhibitor on the susceptibility of peripheral nerves to compression was studied in rats made diabetic by the injection of streptozotocin (50 mg.kg-1). The response to nerve compression was determined in untreated diabetic rats after 22 days of diabetes and compared with the response in two similar groups of diabetic rats which had been treated with the aldose reductase inhibitor 'Statil' (ICI 128436; 25 mg.kg-1.day-1 orally) either from the induction of diabetes or for 7 days prior to nerve compression. Two groups of non-diabetic rats were treated with 'Statil' for either 22 days or 7 days to act as controls. Inhibition of fast axonally transported proteins was induced by local compression of the sciatic nerves 4 h after application of 3H-leucine to the motor neurone cell bodies in the spinal cord. The inhibition of fast axonal transport was quantified by calculation of a transport block ratio. Compression at 30 mmHg for 3 h induced a significantly greater (p less than 0.05) inhibition of axonal transport at the site of compression in nerves of untreated diabetic rats (transport block ratio 0.96 +/- 0.24, n = 8) than in nerves of control rats treated with the aldose reductase inhibitor for either the shorter time of 7 days (0.71 +/- 0.17, n = 10) or the longer time of 22 days (0.69 +/- 0.08, n = 5).(ABSTRACT TRUNCATED AT 250 WORDS)

Morphologic changes in nerve cell bodies induced by experimental graded nerve compression

The effects of experimental nerve fiber compression on the morphology of nerve cell bodies were studied. Rabbit cervical vagus nerves were crushed or subjected to compression at 0 (sham compression), 30, 200, or 400 mm Hg for 2 h. Morphometric measurements and light microscopical evaluation of the nerve cell bodies in the nodose ganglion were carried out 7 days after the injury on the injured and control sides. Crush and compression at 30, 200, or 400 mm Hg induced a slight decrease in total cell profile area compared with the control side, but it was not related to degree of injury. There was a marked decrease in the ratio between nuclear and total cell profile area (nuclear volume density) after compression at 200 and 400 mm Hg, as well as after crush, and to a lesser extent after compression at 30 mm Hg. Compression at 30, 200, or 400 mm Hg as well as crush of the vagus nerve induced migration of the nucleus to the periphery and dispersion of Nissl substance in the cytoplasm of the nerve cell bodies. Sham compression induced no obvious changes in total cell profile area, nuclear volume density, or migration of nucleus. There was a somewhat increased percentage of cells showing dispersion of Nissl substance in sham-compressed animals than in controls. The results show that nerve fiber compression induced pronounced reactive changes in nerve cell bodies, even at low pressures, corresponding to those found in human carpal tunnel syndrome. Such pressures are known to induce reversible inhibition of fast axonal transport as well as inhibition of retrograde axonal transport. The nerve cell body changes in the nodose ganglion may thus be a reaction to disturbances in axonal transport.

Inhibition of fast axonal transport in bullfrog nerves by dibenzazepine and dibenzocycloheptadiene calmodulin inhibitors

The effects of the calmodulin inhibitors amitriptyline, desipramine, imipramine, and clomipramine on fast axonal transport, oxidative metabolism, and density of axonal microtubules were measured in bullfrog spinal nerves in vitro. The four drugs tested inhibited the fast orthograde transport of [3H]leucine-labelled proteins and the fast retrograde transport of acetylcholinesterase at a concentration of 0.2 mM. Amitriptyline, desipramine, and imipramine were equipotent inhibitors of transport, and clomipramine was a more potent inhibitor than imipramine. The adenosine triphosphate content of the nerves was reduced by at most 19% by the compounds under study; such a reduction cannot account for the inhibition of fast axonal transport. Desipramine and imipramine had no significant effect on the density of microtubules in unmyelinated axons, whereas amitriptyline only reduced it by 18%; the inhibition of axonal transport by these three drugs can therefore not be explained by microtubule disruption. Clomipramine reduced microtubular density by 40%, and this effect may have contributed to the inhibition of fast axonal transport. The inhibition of fast axonal transport by desipramine, imipramine, and amitriptyline may be related to the inhibition of calmodulin function by these drugs. The similar potency of these three drugs as inhibitors of fast axonal transport goes in parallel with their known similar potency as calmodulin antagonists.

Effects of nerve compression on fast axonal transport in streptozotocin-induced diabetes mellitus

The hypothesis that nerves in diabetes mellitus exhibit an increased susceptibility to compression was experimentally tested. Inhibition of fast axonal transport was induced by local compression in sciatic nerves of rats with streptozotocin-induced diabetes mellitus. Fast anterograde axonal transport was measured after application of 3H-leucine to the motor neurone cell bodies in the spinal cord. The sciatic nerve was subjected to local, graded compression in vivo by a small compression chamber. The amount of accumulation of proteins was quantified by calculation of a transport block ratio. Compression at 30 mm Hg for 3 h induced a significantly greater (p less than 0.05) accumulation of axonally transported proteins at the site of compression in nerves of diabetic animals (transport block ratio: 1.01 +/- 0.35; n = 7) than in nerves of controls (0.67 +/- 0.16; n = 7). Accumulation was significantly higher in ligature experiments of both control (1.34 +/- 0.44; n = 8; p less than 0.01) and diabetic animals (1.45 +/- 0.30; n = 8; p less than 0.05), indicating that the block of transport in compressed nerves was incomplete. Neither sham compressed diabetic (0.50 +/- 0.09; n = 6) nor control (0.49 +/- 0.11; n = 6) nerves showed any block of axonal transport. The possible causes of the increased inhibition of fast axonal transport in diabetic rats are discussed. The results indicate that diabetes may lead to an increased susceptibility of peripheral nerves to compression.

Inhibition of fast axonal transport by DNase I

Inhibition of fast axonal transport by DNase I